Building Blocks for the Screening of Histone Deacetylase Inhibitors Using μSPOT

Area of Science:

• Biochemistry
• Medicinal Chemistry
• Drug Discovery

Background:

• Histone deacetylase (HDAC) inhibitors are utilized in cancer chemotherapy and investigated for other diseases.
• A significant gap exists in isozyme-selective HDAC inhibitors, hindering the understanding of individual HDAC functions and the development of drugs for broader applications.

Purpose of the Study:

• To synthesize novel hydroxamic acid and trifluoromethyl ketone building blocks.
• To utilize these building blocks for creating diverse libraries of HDAC-binding peptides.
• To establish a method for efficient screening of these peptide libraries for HDAC inhibitory activity and selectivity.

Main Methods:

• Synthesis of hydroxamic acid and trifluoromethyl ketone building blocks.
• Peptide library synthesis using the SPOT method on cellulose discs.
• Deprotection and solubilization to create printable DMSO stocks.
• Screening of peptide libraries on assay-ready slides for HDAC inhibition.

Main Results:

• Successfully synthesized novel building blocks for HDAC inhibitor development.
• Generated diverse peptide libraries with potential HDAC-binding capabilities.
• Demonstrated a method allowing multiple screening rounds on a single library.
• Identified compounds active in cellular assays, exhibiting diverse selectivity.

Conclusions:

• The developed method facilitates the efficient screening of peptide libraries for HDAC inhibitors.
• This approach aids in the discovery of isozyme-selective HDAC inhibitors.
• The findings provide a platform for developing novel therapeutics for a wider range of disorders beyond cancer.